Department of Medicine, Division of Clinical Pharmacology, Room 536 Robinson Research Building, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Department of Pharmacology, College of Pharmacy, University of Sargodha, University Road, Sargodha, Punjab, Pakistan.
Physiol Rep. 2024 Jun;12(11):e16048. doi: 10.14814/phy2.16048.
Studying acute changes in vascular endothelial cells in humans is challenging. We studied ten African American women and used the J-wire technique to isolate vein endothelial cells before and after a four-hour lipid and heparin infusion. Dynamic changes in lipid-induced oxidative stress and inflammatory markers were measured with fluorescence-activated cell sorting. We used the surface markers CD31 and CD144 to identify human endothelial cells. Peripheral blood mononuclear cells isolated from blood were used as a negative control. The participants received galantamine (16 mg/day) for 3 months. We previously demonstrated that galantamine treatment effectively suppresses lipid-induced oxidative stress and inflammation. In this study, we infused lipids to evaluate its potential to increase the activation of endothelial cells, as assessed by the levels of CD54+ endothelial cells and expression of Growth arrest-specific 6 compared to the baseline sample. Further, we aimed to investigate whether lipid infusion led to increased expression of the oxidative stress markers IsoLGs and nitrotyrosine in endothelial cells. This approach will expedite the in vivo identification of novel pathways linked with endothelial cell dysfunction induced by oxidative stress and inflammatory cytokines. This study describes an innovative method to harvest and study human endothelial cells and demonstrates the dynamic changes in oxidative stress and inflammatory markers release induced by lipid infusion.
研究人类血管内皮细胞的急性变化具有挑战性。我们研究了 10 名非裔美国女性，在脂质和肝素输注前和输注后使用 J 型线技术分离静脉内皮细胞。使用荧光激活细胞分选术测量脂质诱导的氧化应激和炎症标志物的动态变化。我们使用表面标志物 CD31 和 CD144 来鉴定人内皮细胞。从血液中分离的外周血单核细胞被用作阴性对照。参与者接受加兰他敏（16mg/天）治疗 3 个月。我们之前证明加兰他敏治疗可有效抑制脂质诱导的氧化应激和炎症。在这项研究中，我们输注脂质以评估其增加内皮细胞激活的潜力，通过 CD54+内皮细胞的水平和与基线样本相比生长停滞特异性 6 的表达来评估。此外，我们旨在研究脂质输注是否导致内皮细胞中氧化应激标志物 IsoLGs 和硝基酪氨酸的表达增加。这种方法将加速鉴定与氧化应激和炎症细胞因子诱导的内皮细胞功能障碍相关的新途径。本研究描述了一种从人内皮细胞中获取和研究的创新方法，并证明了脂质输注诱导的氧化应激和炎症标志物释放的动态变化。
